Characterization of human thyrotropin receptor-related peptide-like immunoreactivity in peripheral blood of Graves' disease. An antiserum raised against an alignment of amino acid-(32-56), termed TSHRP-1, in the extracellular domain of human thyrotropin (TSH) receptor was used to identify the TSH receptor-like substance in plasma of Graves' disease. The dilution curve of plasma TSHRP-1-like immunoreactivity was observed in a manner parallel to the standard synthetic peptide curve in radioimmunoassay, and its molecular weight estimated approximately 60 kDa. The amounts of TSHRP-1-like immunoreactivity were significantly higher in Graves' plasma than those in plasma of normal and hypothyroid patients due to Hashimoto's thyroiditis. The present results indicate that human peripheral blood possesses a soluble form of the extracellular domain of TSH receptor which may contribute to the pathophysiology of Graves' disease.


Phosphorylation of the recombinant spliced variants of the alpha-sub-unit of the stimulatory guanine-nucleotide binding regulatory protein (Gs) by the catalytic sub-unit of protein kinase A. Both GS alpha-1 and GS alpha-4 were phosphorylated by the purified catalytic sub-unit of protein kinase A. Phosphate incorporation into 220 pmol and 190 pmol of GS alpha-4 and GS alpha-1 after a 1 hour incubation with kinase was 14 pmol and 10 pmol, respectively. These low levels of phosphorylation are due to the thermal lability of purified recombinant GS alpha. However, the phosphorylation was inhibited by guanine nucleotides (GDP-beta-S, GppNHp and GTP) and is, therefore, a specific event. We suggest that, as for GS alpha phosphorylation by protein kinase C (Pyne et al., 1992), the guanine nucleotide-free form of GS alpha is the most likely substrate. Guanine-nucleotides reduce the lifetime and, therefore availability for phosphorylation, of guanine-nucleotide free GS alpha. GS alpha phosphorylation by protein kinase A in vitro provides preliminary evidence that a similar phosphorylation of GS alpha may be an important regulatory event in cells.


pH dependent conformational changes in the T- and R-states of insulin in solution: circular dichroic studies in the pH range of 6 to 10. Zinc insulin hexamer has been shown to undergo a phenol-induced T6 to R6 conformational transition in solution. Our circular dichroic (CD) studies demonstrate that insulin undergoes pH-dependent conformational changes over the pH range of 6-10 in the T-state and in the R- state. In order to determine which specific amino acid residues may be responsible for these pH-dependent changes, a series of insulin analogs were utilized. In the T-state, the pH dependent CD changes monitored in the far UV region have a pK of 8.2 and appear to be related to the titration of the A1-Gly amino group. Using the near UV CD a second pH-dependent conformational change was detected with a pK of 7.5 in the T-state. 1H N.M.R. studies suggest that B5-His may be responsible for this conformational transition. In the presence of m-cresol (R-state), the pK value was found to be 6.9. During this titration, the increased ellipticity for the R-state is diminishing as pH decreases from pH 8 to 6, and no difference in ellipticity was observed at 255 nm between T- and R-states at pH 6. Therefore, this may be due to the transition from the R back to the T-state.


Effect of pH on quercetin-induced suppression of heat shock gene expression and thermotolerance development in HT-29 cells. When cells were heated for 15 min at 45 degrees C, they became thermotolerant to a second heat exposure at 45 degrees C. Thermotolerance developed rapidly, reached its maximum 6 hr after heat shock, and then gradually decayed. The development of thermotolerance was partially suppressed by treatment with various concentrations of quercetin (0.05-0.2 mM) at pH 7.4 after the initial heat treatment. In contrast, the drug markedly inhibited thermotolerance development at pH 6.5. Furthermore, a combination of low pH and quercetin treatment distinctively altered the expression of HSP70 gene compared with that of HSP28 or HSP90 gene. These results demonstrate a good correlation between the amount of HSP70 gene expression and development of thermotolerance.


Detection of the GLUT3 facilitative glucose transporter in rat L6 muscle cells: regulation by cellular differentiation, insulin and insulin-like growth factor-I. The GLUT3 facilitative glucose transporter protein was found to be expressed in rat L6 muscle cells. It was detected at both the myoblast and myotube stage. GLUT3 protein content per mg of total membrane protein increased significantly during L6 cell differentiation. Subcellular fractionation demonstrated that the GLUT3 protein was predominantly localized in plasma membrane-enriched fractions of either myoblasts or myotubes. Short-term exposure of L6 myotubes to IGF-I or insulin caused a redistribution of GLUT3 protein from an intracellular membrane fraction to the plasma membrane, without affecting total membrane GLUT3 protein content. Long-term exposure of L6 myotubes to IGF-I produced an increase of GLUT3 protein in total membranes and all subcellular membrane fractions, especially the plasma membrane. We propose that the GLUT3 glucose transporter may play an important role in glucose metabolism in developing muscle.


Rhamnose moiety of phosphoramidon is not required for in vivo inhibition of endothelin converting enzyme. Experiments were conducted to determine the importance of the carbohydrate moiety of phosphoramidon in the inhibition of the pressor response to big endothelin-1 in anesthetized rats. Big endothelin-1 produced a 42% increase in mean arterial pressure which was nearly abolished by co-infusion of phosphoramidon. Similarly, when an analog of phosphoramidon lacking the rhamnose group (phosphoryl-leu-trp-OH) was co-infused, a significant attenuation of the pressor response was observed. These findings indicate that the rhamnose moiety of phosphoramidon is not responsible for the distinguishing this compound as an inhibitor of the response to big endothelin-1 in the rat.


High level expression, purification, and characterization of the Kunitz-type protease inhibitor domain of protease nexin-2/amyloid beta-protein precursor. The protease inhibitor, protease nexin-2 (PN-2), is the secreted isoform of the Alzheimer's amyloid beta-protein precursor (A beta PP) that contains the Kunitz-type protease inhibitor (KPI) domain. Here we describe the use of the methylotrophic industrial yeast Pichia pastoris as a host system for the large scale production of the KPI domain of PN-2/A beta PP. In addition to the 57 amino acid KPI domain, the expression product contained an additional four amino acid residues at its amino terminus that correspond to amino acids 285-288 of A beta PP (Ponte et al. 1988 Nature 311:525-527). This expression system generated yields of greater than 1.0 gram of KPI domain per liter of fermentation media. The secreted 61 amino acid product was purified to homogeneity and biochemically characterized. Amino acid analysis and sequencing of the entire expressed KPI domain verified its integrity. Similar to native PN-2/A beta PP, the purified KPI domain potently inhibited trypsin, chymotrypsin, and coagulation factor XIa. Although heparin augments the inhibition of factor XIa by native PN-2/A beta PP it had no effect on the inhibition of factor XIa by expressed KPI domain suggesting that heparin binds to regions on native PN-2/A beta PP outside of the protease inhibitory domain. This KPI domain expression product should be useful in studying the physiologic and pathophysiologic functions of PN-2/A beta PP.


The generation of inositolglycan mediators from rat liver plasma membranes: the role of guanine nucleotide binding proteins. The guanine nucleotide dependence for the generation of inositolglycan second messengers from rat liver plasma membranes has been investigated. Plasma membranes, when treated with insulin release a soluble mediator substance which activates pyruvate dehydrogenase (PDH). Guanosine 5'-[3-thio]triphosphate (GTP gamma S) was found to be as potent as insulin in stimulating mediator release. The stimulatory effects of GTP gamma S required the presence of magnesium and following preincubation of membranes with guanosine 5'-[2-thio]diphosphate (GDP beta S) the stimulation of mediator release by either insulin or GTP gamma S was blocked. The activation of PDH by mediator fractions produced in response to either insulin or GTP gamma S was abolished following treatment of the fractions with anti-inositolglycan antibodies. The significance of these observations with respect to the possible involvement of a regulatory guanine-nucleotide binding protein (G-protein) in the generation of insulin mediators is discussed.


Correlation between up-regulation of lymphokine mRNA and down-regulation of TcR, CD4, CD8 and lck mRNA as shown by the effect of CsA on activated T lymphocytes. Full activation of different T cell populations via the TcR/CD3 complex leads to transient expression of lymphokine mRNA and to transient and specific down-modulation of TcR, CD4, CD8 and p56lck mRNA (Multi-Receptor Complex or MRC mRNA). This transient down-modulation is due to both a decrease in transcription and stability of these mRNA and is temporally and quantitatively related to lymphokine mRNA induction. We showed that cyclosporin A (CsA), which blocks lymphokine expression also inhibits MRC mRNA down-modulation at the transcriptional level, and does not affect mRNA stability. The fact that CsA inhibits both lymphokine expression and MRC mRNA down-modulation at transcriptional level supports a model in which similar signals trigger the inverse regulation of these two sets of genes via identical transcriptional factors.


Hydrogen/deuterium isotope effect in the oscillating peroxidase-oxidase reaction. The oscillatory peroxidase-oxidase reaction has been investigated by using NADH deuterated in the nicotinamide 4-A position. A considerable kinetic hydrogen/deuterium isotope effect on the oscillatory behavior was revealed, which may provide an additional valuable tool for mechanistic studies and for discriminating between various mechanistic models of the peroxidase-oxidase reaction. Particularly, this effect manifests in different oscillation frequencies. A sequence of simple and aperiodic oscillations was found between two stable steady states.


Glycation of lens MIP26 affects the permeability in reconstituted liposomes. We studied the role of glycation of lens putative gap junctional protein, MIP26, on the permeability as well as on calmodulin mediated gating activity in reconstituted liposomes. Calf lens membranes were incubated with 0-100 mM glucose for 3 days and MIP26 was isolated. There was a glucose concentration dependent increase in the glycation of MIP26 which reached to 2.48 moles/mole of protein with 100 mM glucose. Gel electrophoresis showed that there was no degradation of MIP26 to MIP22 during incubation. Channel permeability was determined by reconstituting MIP26 into asolectin liposomes. There was a MIP26 glycation dependent decrease in the permeability to sucrose. Furthermore, proteoliposomes containing nonglycated MIP26 showed complete uncoupling of the channels with calmodulin whereas the channels containing glycated MIP26 were only partially uncoupled. These results suggest that glycation of MIP26 does interfere with the gating activity in reconstituted liposomes.


Proteolytic machinery of glomerular epithelial cells against IgG. Accumulation of immune complexes in the subepithelial region of the glomerular basement membrane results in the lesion of membranous nephropathy. The inefficient handling of immune complexes by the glomerular epithelial cell was investigated by studying the mechanism of IgG proteolysis by the intracellular proteases of cultured epithelial cells. Radiolabelled IgG was incubated with extracts of cells and the digestion of IgG was monitored by SDS-PAGE analysis. Prolonged incubation of IgG with the cell extracts resulted in only partial degradation of the IgG. The enzyme responsible for the breakdown was determined to be the lysosomal cathepsin D based on the pH optimum and the presence of aspartate in the active site of the enzyme. SDS-PAGE analysis of the digestion fragments revealed that a large proportion of the incompletely degraded IgG was the (Fab)2 fragment, which was resistant to further proteolysis. This could be one of the possible explanations for the slow removal of IgG from the subepithelial location of the basement membrane.


Low-affinity Ca(2+)-binding sites versus Zn(2+)-binding sites in histidine-rich Ca(2+)-binding protein of skeletal muscle sarcoplasmic reticulum. Histidine-rich Ca(2+)-binding protein (HRC) is a 170 kDa protein that can be identified in the isolated sarcoplasmic reticulum from rabbit skeletal muscle by its ability to bind [125I]low-density lipoprotein on blots after SDS-PAGE and that appears to be bound to the junctional membrane through calcium bridges. Molecular cDNA cloning of this protein predicts the existence of a Ca(2+)-binding domain and of a distinct heavy-metal binding domain at the cystein-rich COOH-terminus. Here we demonstrate, using radioactive ligand blot techniques, that HRC protein binds 45Ca at low affinity, as well as being able to bind 65Zn, but at different sites, that are largely inhibitable by prior reductive alkylation of the protein. In contrast to Ca(2+)-binding protein calsequestrin not having detectable 65Zn-binding sites, HRC protein bound selectively to immobilized Zn2+ on IDA-agarose affinity columns. Our results also indicate that rabbit and human 140 kDa HRC protein have common properties.


Calreticulin is present in the acrosome of spermatids of rat testis. We have already succeeded in purifying a calcium-binding protein (CalBP) from rat spermatogenic cells [Nakamura et al., Biochem. Biophys. Res. Commun., 176 (1991) 1358]. In this study, the location of this protein within rat testis was examined, using a rabbit antisera for this protein. The antigen was localized on the developing acrosomes during spermiogenesis. The NH2-terminal amino acid sequence obtained for rat CalBP was identical to that of calreticulin obtained for the skeletal muscle of mice and closely resembled that for rabbit calreticulin. On the immunoblot analysis, the purified rat CalBP reacted with an antibody raised against rabbit skeletal muscle calreticulin. The results indicate that calreticulin is present in the acrosome of spermatids of rat testes.


The regulation of the intracellular sodium ion concentration in cultured chick embryo heart cells. Using digital imaging microscopy with the fluorescent indicator sodium-binding benzofuran isophtalate, we examined the cytosolic Na+ concentration ([Na+]i) in individual chick embryo heart cells. Inhibition of the Na(+)-H+ exchanger using Na(+)-free (Li+ substituted) medium and inhibition of the Na(+)-efflux through the Na(+)-Ca2+ exchanger using Ca(2+)-free medium didn't change the [Na+]i. The opening of voltage-dependent Na+ channels with veratridine (150 micrograms/ml) and inhibition of the Na(+)-K(+)-Cl(-)-cotransporter with bumetanide (10 microM) led to an increase in [Na+]i by 107% and 86%, respectively, suggesting that the Na+ channels and the Na(+)-K(+)-Cl- cotransporter predominantly regulate the [Na+]i in cultured chick embryo heart cells.


Identification of the iron-binding histidine residues in soybean lipoxygenase L-1. Lipoxygenases constitute a class of non-heme, non-sulfur iron dioxygenases acting upon lipids possessing a 1,4-cis-cis-pentadiene moiety. The iron is known to be essential for activity. A motif of six histidine residues has been found in all of the thirteen lipoxygenases, from both plant and animal sources, whose sequences have been reported. We had previously obtained mutant proteins in which each of the 6 conserved histidines of soybean lipoxygenase L-1 had been replaced and found that the mutants H499Q, H504Q (or H504S) and H690Q had no detectable enzymatic activity. We have now found that these inactive proteins contain no Fe, although they have the same electrophoretic mobility as wild-type L-1 under both denaturing and non-denaturing conditions and react with anti-L-1 antibodies.


Quinone dependent NADH dehydrogenation in mitochondria-like particles from Setaria digitata, a filarial parasite. In the cattle filarial parasite, Setaria digitata, the mitochondria-like particles have been shown to possess site I associated oxidative phosphorylation and rotenone sensitive and insensitive pathways for the dehydrogenation of NADH. Quinone depleted mitochondria-like particles show a loss of activity of these NADH dehydrogenases and also a complete loss of fumarate reductase activity. Reconstitution with quinone restores both NADH linked oxygen uptake and fumarate reductase activity. Thus activities of complex I and fumarate reductase are linked to quinone. Hence an inhibitor at the level of quinone can simultaneously block both aerobic and anaerobic pathways which drive ATP production and may prove useful in the effective control of filariasis.


Transactivation of the human c-myc gene by c-Myb. We analyse the contribution of six Myb-binding sites in the upstream c-myc sequences to transactivation by co-transfection assays. Surprisingly, deletion of the six Myb-binding sites did not influence the transactivation of c-myc by c-Myb protein. Instead, the strongest transactivation was observed with a c-myc reporter plasmid which contains only 450 bp of exon 1 including the c-myc promoter P2. An exchange of the DNA binding domain of c-Myb by that of GAL4 led only to small transactivation effects indicating that the DNA binding domain of c-Myb is essential for transactivation of the c-myc gene. These results suggest either an indirect transactivation mechanism of the c-myc gene by c-Myb proteins or a role of the DNA binding domain for additional effects than DNA binding.


Identification of four genomic loci highly related to casein-kinase-2-alpha cDNA and characterization of a casein kinase-2-alpha pseudogene within the mouse genome. Using the coding region of the human CK-2 alpha cDNA as a probe for screening a genomic mouse library, positive clones representing four different genomic loci were isolated. Partial DNA sequences of these loci encompassing the first 120 nucleotides of the putative coding region are reported. One positive clone was further analyzed by sequencing a 3.1 kb XbaI fragment. This clone displays the characteristics of a pseudogene, i.e. lack of introns and several nucleotide insertions and deletions. In its 3' region it contains a 91 bp large CT-rich stretch which consists of (CCTT) and (CT) repeats; in the 5' region three (CCCCCT) repeats.


Purification and properties of a functional 47-kilodalton cytosolic factor required for NADPH-oxidase activation in bovine neutrophils. A cytosolic factor of 47 kDa required for activation of the NADPH oxidase, and referred to as p47, has been purified in its functional form from the cytosol of resting bovine neutrophils. The purification was monitored by the determination of the activating potency of p47 in a cell-free system of oxidase activation. The recovery was around 10% and the purification factor greater than 1000. P47 was phosphorylated in vitro by protein kinase A and protein kinase C. [32P] labeled p47 was resolved by isoelectric focusing into two major labeled bands of pI 7.0 and 8.5. Polyclonal antibodies were used to demonstrate that p47 is localized specifically in the cytosol of resting neutrophils, and that, upon activation of neutrophils, p47 is translocated from the cytosol to the membrane.


Functional characterisation of serum DNase I in MRL-lpr/lpr mice. The autosomal defect in Fas antigen leads CD4-CD8-T-cells to accumulate in lymph nodes and spleen of MRL-lpr/lpr mice. MRL-lpr/lpr mice present increased levels of DNase I as compared to the control strain MRL-+/+. This DNase I, which most probably originates from the accumulated CD4-CD8-T-cells, cleaves nuclear DNA with a strong preference for internucleosomal sites yielding, in the presence of both Ca2+ and Mg2+, a pattern of fragments typical for apoptosis. Furthermore, we show that this "apoptosis-ladder" can be obtained with purified DNase I in presence of normal serum.


A mathematical approach to lipolysis based on the interrelationship of physicochemical and biochemical data. A novel approach is described for the quantitation of the parameters involved in lipolytic activity. The activity of lipase purified from human pancreatic juice against an oil-water emulsion depends on the degree of emulsification, the type of emulsifier used, the type and the concentration of bile salt, the concentration of colipase and, finally, on the zeta-potential of the interface. The zeta-potential, in turn, depends on the partition of bile salt molecules between the micellar state in the bulk and their adsorption on the oil-water interface. None of these parameters correlates straightforward with lipase activity. However, if several parameters are taken into account, the enzyme activity determined in our system obeys a multiparametric equation where the variables are: the conjugation of bile salts, the number of hydroxy groups, their aggregation number and, finally, the zeta-potential of the oil droplets.


Human preproendothelin-1 is converted into active endothelin-1 by baculovirus-infected insect cells. To investigate biochemical and biological parameters involved in preproendothelin-1 (preproET-1) maturation we infected Spodoptera frugiperda (Sf21) cells with a suitable engineered baculovirus vector carrying the cDNA encoding the entire human 212 amino acids precursor. Culture supernatants were tested by RIA using an anti-ET-1 serum, ET-1-like immunoreactive material (IRM) was detected in the infected Sf21 cells but not in control, wild-type or mock-infected cells. Fractionation of the culture supernatant by RP-HPLC coupled to an ET-1 specific RIA yielded two main peaks corresponding to the retention times of human bigET-1 and ET-1. Furthermore, culture supernatant of preproET-1 expressing Sf21 cells elicited a characteristic dose-response vasoconstrictive activity on rabbit vena cava, consistent with the amount of ET-1 as estimated by RP-HPLC coupled to RIA. These results suggest that insect cells possess the enzymatic activities necessary for human preproET-1 full maturation even though no such peptide has ever been found in insect cells.


Evaluation of nitrite production by human monocyte-derived macrophages. Reactive nitrogen intermediates are important in the anti-tumor and anti-microbial activities of rodent macrophages, but it is not known whether this is the case for human macrophages. In the present study, nitrite concentrations in vitro were used as an indicator of reactive nitrogen intermediate production by mouse, rat, and human macrophages. Human macrophages derived by culturing peripheral blood monocytes did not consistently produce detectable nitrite levels in response to any stimulus examined. Human macrophages were viable and metabolically active as indicated by the MTT assay, and their respiratory burst response to phorbol myristate acetate was increased following incubation with Interferon-gamma, as expected for typical macrophages. In contrast, rat or mouse peritoneal macrophages produced nitrite concentrations of approximately 20-100 microM in response to lipopolysaccharide, Interferon-gamma, or both. These results demonstrate substantial differences in the production of nitrites by rodent and human macrophages. Because of the heterogeneity among macrophage populations, these findings may not be applicable to all human macrophage populations, but they suggest a need for caution in extrapolating from rodent studies regarding the role of reactive nitrogen intermediates in anti-tumor or anti-microbial functions of human macrophages.


Purified estrogen receptor enhances in vitro transcription. An in vitro transcription system was developed to investigate the mechanisms of gene regulation by the estrogen receptor (ER). ER purified from calf uterus was highly active in enhancing RNA transcription from a template DNA containing estrogen response elements (EREs) upstream from a minimal promoter. Under the conditions employed, no addition of tissue specific factors was required and both estrogen or antiestrogens were ineffective. The stimulation of transcription correlated with the copy number of EREs in the template. The addition of competitor ERE oligonucleotides specifically inhibited the ER-induced transcription. We suggest that the ER may be involved in the formation of the stable initiation complex.


Signal peptide for peroxisomal targeting: replacement of an essential histidine residue by certain amino acids converts the amino-terminal presequence of peroxisomal 3-ketoacyl-CoA thiolase to a mitochondrial signal peptide. The role of the histidine residue at position -17 of the amino-terminal signal peptide of rat peroxisomal 3-ketoacyl-CoA thiolase was studied in vivo, employing site-directed mutagenesis. Among the nine amino acids tested, only glutamine could partially substitute for the histidine. Mutants carrying basic amino acids, arginine and lysine, and hydrophobic residues, leucine and valine, in place of histidine were all translocated to mitochondria, but not to peroxisomes. These results indicate that the signal peptide of the thiolase is recognized by a mechanism totally different from that for the SKL motif, a known peroxisomal targeting signal. Relationship of the thiolase signal peptide to those of mitochondrial proteins is discussed.


Isolation and sequence determination of cDNA encoding PMP-22 (PAS-II/SR13/Gas-3) of human peripheral myelin. A full length cDNA of PMP-22 (PAS-II/SR13/Gas-3) of peripheral myelin has been isolated from a cDNA library of human fetus spinal cord. The clone is 1823 base pairs (bp) in length and contains a 480 bp open reading frame encoding a polypeptide of 160 residues. The deduced amino acid sequence is highly homologous to PMP-22 from bovine (PAS-II), rat (SR13) and mouse (Gas-3).


Protein transport in the permeabilized cell of Schizosaccharomyces pombe. We reconstituted a protein translocation-transport system composed of permeabilized spheroplasts (P-cells) of the fission yeast Schizosaccharomyces pombe and the precursor of alpha sex pheromone, prepro-alpha-factor of the budding yeast Saccharomyces cerevisiae. We found that P-cells prepared from the spheroplasts formed in 0.7M KCl as an osmotic stabilizer had the activity to transport pro-alpha-factor to the Golgi apparatus. Electron microscopic observations showed that membranes were preserved more intact in the P-cells prepared from the spheroplasts formed in 0.7M KCl than in 0.7M sorbitol. A glycoprotein of S. pombe contains galactose residues, and we detected incorporation of radiolabeled galactose residues into the anti-prepro-alpha-factor immunoprecipitable fractions in this S. pombe system, but not in the S. cerevisiae system. This paper reports that a heterologous system of in vitro protein transport was performed, and prepro-alpha-factor has the signals necessary for early steps of the transport in S. pombe.


In vivo targeting of inflamed areas by electroloaded neutrophils. Due to their spontaneous accumulation in inflamed or infected areas, blood phagocytes are potent drug vectors with specific targeting. Drug like molecule loading was obtained by use of cell electropermeabilization in which the impermeability of their plasma membrane is transiently impaired. Electrical conditions were used which allow electroloading of a drug like molecule (propidium iodide) in 70% of leukocytes in a whole blood sample while preserving in vitro functional properties. Slow release of entrapped hydrophilic molecules was observed with a half lifetime longer than 4 hours at 4 degrees C and at 37 degrees C. With an in vivo assay, using a rat model of inflammation, we showed that, as for non-pulsed cells, pulsed neutrophils accumulate 10 times more in an inflamed area than they do in control areas. Phagocyte electropermeabilization is therefore a very efficient way of drug targeting. Accumulation of electropulsed neutrophils in an area of inflammation gives targeted release of the electroloaded drug.


Kinetic analysis of duck epsilon-crystallin with L-lactate dehydrogenase activity: determination of kinetic constants and comparison of substrate specificity. A systematic analysis of the kinetic properties of duck lens epsilon-crystallin with lactate dehydrogenase [LDH, (E.C. 1.1.1.27)] activity was carried out by employing some 19 different alpha-keto acids as substrates for this NADH-dependent LDH-catalyzed reaction. The steady-state Michaelis and catalytic constants (Km, kcat) were determined for a broad range of organic compounds. The results provide important insights regarding the binding and affinity of substrates to active sites of this enzyme crystallin and indicate a great potential for the application of the stable epsilon-crystallin as a catalyst to the synthesis of some important chiral alpha-hydroxyacids in a convenient and efficient way. It is also demonstrated for the first time that in addition to the enzymatic activity of lactate dehydrogenase, duck epsilon-crystallin also possesses the enzymatic activity of malate dehydrogenase.


Homotypic and heterotypic Ca(++)-independent cell adhesion activities of biliary glycoprotein, a member of carcinoembryonic antigen family, expressed on CHO cell surface. Homotypic and heterotypic cell adhesion activities of a carcinoembryonic antigen (CEA) family member, biliary glycoprotein a (BGPa), have been examined. CHO cells transfected with the cDNA for BGPa, CEA, non-specific cross-reacting antigen (NCA) and CGM6 have been used. The BGPa producers showed both homotypic and heterotypic adhesion to CEA and NCA producers. However, they hardly adhered to CGM6 producers. Calcium ion was not required for BGPa-mediated homotypic and heterotypic cell adhesion as well as for the adhesions of other members of CEA family. The results strongly suggested that BGPa may play some important roles through Ca(++)-independent cell adhesion activities.


Regulatory role of microsomal 3-hydroxy-3-methylglutaryl-coenzyme A reductase in a tobacco mutant that overproduces sterols. In a tobacco mutant callus, containing up to tenfold more sterols than the wild-type genotype, HMG-CoA reductase activity is increased by a factor of approximately three, as is the case in mutant seedlings and plants. The rate of HMG-CoA synthesis from acetyl-CoA by the coupled enzyme system acetoacetyl-CoA thiolase/HMG-CoA synthase, as well as its conversion to acetyl-CoA plus acetoacetate by action of HMG-CoA lyase are not affected. These results confirm the key-regulating role of HMG-CoA reductase in sterol biosynthesis, which seems not to be confined only to the animal kingdom, but can also be extended to plants.


A homology domain shared between Drosophila optomotor-blind and mouse Brachyury is involved in DNA binding. The distribution of sequence elements divides the optomotor-blind protein into three regions and is suggestive of a transcriptional regulatory role of this protein. The central region of Omb is homologous to the N-terminal half of the Brachyury protein. The conserved domain of Omb is here shown to possess general DNA binding affinity but has no significant similarity to recognized DNA binding motifs.


Mapping of the dopa decarboxylase gene to the 11A band of the murine genome. A human DOPA decarboxylase (DDC) cDNA probe of 747 base pairs has been used to map the DDC gene by in situ hybridization on mouse metaphase chromosomes. This result indicates that the gene is located on band 11A, near the erythroblastosis oncogene B (erb b) locus. This provides evidence for a synteny group on mouse chromosome 11 and human chromosome 7.


13C NMR spectroscopic measurement of glutathione synthesis and antioxidant metabolism in the intact ocular lens. A 13C NMR spectroscopic method for non-invasive, time-resolved measurements of glutathione function in the intact ocular lens maintained in organ culture is described. L-[beta-13C]cysteine (1 mM) included in the incubation medium is incorporated, by way of lenticular amino acid uptake and glutathione biosynthetic mechanisms, into the cysteinyl residue of intralenticular glutathione. 13C-NMR chemical shift measurements facilitate analysis of glutathione synthesis and anti-oxidant reactions in the intact tissue. The results of this preliminary study demonstrate the viability of a rapid non-invasive method for monitoring the multiple aspects of glutathione biosynthesis, metabolism, and function in intact tissue.


Vitamin E protects nerve cells from amyloid beta protein toxicity. The amyloid beta protein (ABP) is a 40 to 42 amino acid peptide which accumulates in Alzheimer's disease plaques. It has been demonstrated that this peptide and a fragment derived from it are cytotoxic for cultured cortical nerve cells. It is shown here that ABP and an internal fragment encompassing residues 25 to 35 (beta 25-35) are cytotoxic to a clone of PC12 cells at concentrations above 1 x 10(-9)M and to several other cell lines at higher concentrations. Between 10(-9) and 10(-11) M beta 25-35 protects PC12 cells from glutamate toxicity. The antioxidant and free radical scavenger vitamin E inhibits ABP induced cell death. These results have implications regarding the prevention and treatment of Alzheimer's disease.


Ceruloplasmin stimulates NADH oxidation of pig liver plasma membrane. NADH oxidation by pig liver plasma membranes is stimulated by ceruloplasmin (CUP) reaching a maximal value at 50 U/ml of CUP. NADH oxidation activated by CUP is proportional to the amount of protein. Concanavalin A (Con A) which recognizes the glucidic residues of the CUP required for binding to the receptor inhibits the NADH oxidation in a dose-responsive manner. Both adriamycin and bathophenantroline disulfonate (BPS), previously reported as transplasma membrane electron transport inhibitors, also inhibit the CUP-stimulated NADH oxidation of pig liver plasma membranes. Our results show a clear interaction between CUP and the NADH oxidase of plasma membrane, which supports an oxidative role for CUP in its growth effect.


Footprinting evidence for close contacts of the yeast tRNA(Asp) anticodon region with aspartyl-tRNA synthetase. Chemical footprinting experiments on brewer's yeast tRNA(Asp) complexed to its cognate aspartyl-tRNA synthetase are reported: they demonstrate that bases of the anticodon loop, including the anticodon itself, are in close proximity with the synthetase. Contacts were determined using dimethylsulfate as the probe for testing reactivity of guanine and cytosine residues in free and complexed tRNA. Results correlate with the decrease in aspartylation activity of yeast tRNA(Asp) molecules mutated at these contact positions and will be compared with other structural data arising from solution and crystallographic studies on the aspartic acid complex.


Phorbol ester-mediated suppression of cytochrome P450 Cyp1a-1 induction in murine skin: involvement of protein kinase C. Epidermal 7-ethoxyresorufin O-deethylase (EROD) activity was elevated greater than 100-fold within 4 to 7 h of topical treatment of SENCAR mice with 100 nmol dibenz[a,c]anthracene (DB[a,c]A). Treatment of skin with 2 micrograms of 12-O-tetradecanoylphorbol-13-acetate (TPA) 2 to 8 h prior to DB[a,c]A application suppressed induction by 80%. Suppression was dose-dependent over the range of 0.01 to 5 micrograms TPA (ID50 approximately 0.6 nmol). EROD activities in normal and TPA-treated epidermis paralleled steady state P450 CYP1A1 mRNA content. Analogs of TPA incapable of activating or down-regulating protein kinase C (PKC) did not suppress induction. Pretreatment of skin with sn-1,2-didecanoylglycerol, an activator of PKC which causes translocation but no down-regulation, did not suppress EROD induction. However, induction was suppressed by chrysarobin, an anthralin analog that causes PKC down-regulation in the absence of prior activation. These studies suggest that PKC participates in the processes associated with Cyp1a-1 induction and that TPA effects Cyp1a-1 induction through its down-regulation of PKC.


Antisera to calreticulin inhibits sperm motility in mice. Mouse sperm were rapidly immobilized when exposed to rabbit antisera against rat calreticulin. The inhibition of sperm motility was concentration dependent at dilutions 1:50-350. The velocity of sperm did not change significantly as long as they were motile. Neither motility nor velocity of sperm was affected by adding sheep antisera to bovine calmodulin. The antisera to calreticulin also inhibited in vitro fertilization of mouse eggs. Using the avidin-biotin-peroxidase complex method, the antigen was found to be localized in the acrosome of sperm. The results indicate that calreticulin is present in the acrosome of mouse sperm and may play an important role in sperm motility.


Relaxin increases rat heart rate by a direct action on the cardiac atrium. Relaxin (Rlx) is best understood as a protein hormone of pregnancy that can influence pelvic and cervical connective tissue as well as uterine smooth muscle activity. Thus, it was unexpected that dense Rlx binding sites would be found in the rat cardiac atrium. To functionally characterize this finding, isolated rat atria were challenged with Rlx (0.3 to 30 ng/ml), and they responded with an increased rate (+36%) and force (+38%) of contraction Further studies in conscious normotensive and spontaneously hypertensive rats established by minipump circulating Rlx levels of about 0.5 and 5 ng/ml over 1 to 2 wks. There were significant increases in heart rate of 10-15%, with no consistent changes in blood or urine volume, water or food intake, and mean arterial pressure. We conclude that Rlx can directly stimulate rat cardiac atrial activity in vitro and cause chronotropy in vivo.


Synthesis and pharmacological evaluation of some phenothiazines as antidepressants. 10-Hydrazino acetyl phenothiazine (II) has been converted to N-10-acetyl amino phenothiazine-N-phenyl thiourea (III) which have been converted to 3-aryl-1-(10-phenothiazine acetyl amino)-2,3-dihydro-2-thioxo-4,6-(1H,5H) pyrimidinones (IV) on condensation with aryl amines and aryl aldehydes yielded 3-aryl-1-(10-Phenothiazine acetyl amino)-5-(substituted phenyl amino methyl)-2,3-dihydro-2-thioxo-4,6-(1H, 5H) pyrimidinediones (Va-k) and 3-aryl-1-(10-phenothiazine acetyl amino)-5-(substituted phenylidine)-2,3-dihydro-2-thioxo-4,6-(1H, 5H) pyrimidinediones (vl-o). The compounds were screened for their antidepressant activity against a tricyclic antidepressant (imipramine). Compounds Va, Vf, Vm and Vi exhibited activity better than imipramine with no toxicity (ALD50 greater than 1000 mg/kg) but Vi showed some side effects.


Effect of vinpocetine on red blood cell deformability in vivo measured by a new centrifugation method. A simple new apparatus has been devised to measure red blood cell deformability and the effect of vinpocetine (CAS 42971-09-5) on this property was examined in vivo. The filterability index obtained by this new method correlated well with that obtained by Swank's screen filtration pressure apparatus. Vinpocetine, given as a single oral dose of 10 mg, increased the red blood cell deformability of 5 healthy male volunteers as determined by both methods. The new centrifugation method is simple and suitable for clinical application. Vinpocetine appears to offer a new treatment of cerebral circulatory disorders.


Steady state concentrations and diurnal fluctuations of carbamazepine in patients after different slow release formulations. The bioavailability and serum level fluctuations of three carbamazepine (CBZ, CAS 298-46-4) slow release preparations marketed in Germany were compared in patients with epilepsy. Ten patients who were on CBZ monotherapy and had reached a steady state with morning trough levels above 10 micrograms/ml received all 3 preparations in sequential 3-day periods. On the 3rd day of each period, a 24-h serum drug level profile was determined. Only minimal differences could be found. One preparation had a significantly higher AUC than one of the others (p less than 0.05), and this difference seemed to be correlated with higher fluctuations of serum levels and, especially, an increased Cmax. Although the group differences are minimal, more important differences could be observed individually. One patient had, with one of the three preparations, a reduction in seizure frequency but simultaneously developed important signs and symptoms of toxicity. It is concluded that all 3 preparations are equally suitable for chronic antiepileptic drug treatment. In a well-controlled patient, however, interchange between preparations is not advisable as their kinetics in individual patients can be sufficiently different to cause toxicity or seizure relapse.


Negative inotropic effects of the new class I antiarrhythmic agents berlafenone and alprafenone on electrically stimulated isolated cardiomyocytes. Isolated cardiac myocytes from adult rats were used in a stimulation chamber to investigate the negative inotropic effects of propafenone and the new compounds berlafenone (1-2'-biphenyloxy)-3-tert-butylamino-propanol-2-hydrochloride, GK 23 G; CAS 18965-97-4) and alprafenone (1-(4-methylphenyl)-3-[3'-(2-hydroxy-3-tert-pentylaminopropoxy)-4'- methoxyphenyl]-1-propanonhydrochloride, AH 141; CAS 124316-02-5). This chamber is part of a new device that allows the simultaneous evaluation of mechanics and of the energetics of electrically induced contractions of the myocytes. 1. The contractile behaviour of attaches myocytes was analysed by an image processing system using digitized frames of a CCD camera. 2. The metabolic demand for excitation-contraction coupling was calculated from the drop in oxygen tension (registered by a Clark electrode) caused by suspended myocytes when stimulated in the presence of the contraction inhibiting agent 2,3-butanedione monoxime in the stimulation chamber. 3. The apparent refractory period was evaluated by pacing the myocytes with increasing stimulation rates and determining the frequency at which failure of single contractions occurred. All 3 agents produced a reduction in contraction amplitude of the electrically stimulated myocytes with a similar dose-response relationship (IC50 approx. 10 mumol/l). 4 mumol/l berlafenone reduced the contraction amplitude to 62% of control. Under these conditions the energy expenditure of the contracting cells for excitation and excitation-contraction coupling (ion-cycling) was also reduced (77 +/- 17% of control). Since most of this energy is used for Ca2+ (greater than 80%) it may be concluded that a reduced Ca2+ release causes the negative inotropic action of berlafenone.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of magnesium pyridoxal 5-phosphate glutamate on the hamster cardiomyopathy. 30-day-old polymyopathic hamsters (strain BIO 82.62) were orally treated with either magnesium pyridoxal 5-phosphate glutamate 100 mg/kg, equimolar amounts of magnesium alone (MgCl2 55 mg/kg), or water b.i.d. for 30 days, or sacrificed before treatment. 60-day-old healthy hamsters (strain CLAC) served as controls. Magnesium pyridoxal 5-phosphate glutamate lowered the myocardial calcium content (42.8 +/- 13.1 mmol/kg dry weight) as compared to MgCl2 (70.3 +/- 11.9 mmol/kg dry weight) and water (72.7 +/- 13.6 mmol/kg dry weight). The serum and tissue lipid pattern did not differ between BIO 82.62 and normal hamsters except high serum cholesterol and triglyceride levels in 30-day-old BIO 82.62 hamsters. Myocardial necroses were influenced by neither magnesium pyridoxal 5-phosphate glutamate nor MgCl2. Cytochemical investigation of the ultrastructural calcium localization in the aorta revealed less calcium precipitates with magenesium pyridoxal 5-phosphate glutamate in media myocytes. It is concluded that magnesium pyridoxal 5-phosphate glutamate exerts a calcium antagonistic effect in the cardiomyopathy of the Syrian hamster which is related neither to the magnesium content of magnesium pyridoxal 5-phosphate glutamate nor to its lipid lowering activity.


[Absolute bioavailability of beta-acetyldigoxin from tablets and drops in healthy subjects]. In a crossover design in random order 12 healthy male volunteers were given either beta-acetyldigoxin (Novodigal, CAS 5511-98-8) tablets, oral solution or i.v. application at a digoxin equivalent dose of 0.284 mg. To reach steady state each preparation was given for 10 days on a once-daily schedule. On days 8, 9 and 10 of each observation period blood was sampled to determine trough concentrations of digoxin in steady state. In addition, on day 10 blood was collected repeatedly at appropriate time intervals and urine was sampled concomitantly for 24 h. Trough values during steady state and 24 h AUC were used to calculate digoxin bioavailability for tablets and oral solution. From trough values, the mean bioavailability for beta-acetyldigoxin tablets was 91.2% (range 73.1-118.1) and for solution 93.8% (range 65.7-114.8). Using the AUCs 0-24 h at steady state bioavailability was calculated 77.7% for the tablets and 84.5% for the solution. Since trough values in steady state represent the body burden of digoxin which is supposed responsible for the therapeutic effect, trough values should be given priority for the determination of digoxin bioavailability from beta-acetyldigoxin tablets and solution. All formulations were well tolerated. No clinically relevant side effects were observed.


Influence of isosorbide-5-mononitrate on the circadian rhythm of angina pectoris. Efficacy and safety of two different dose regimens of isosorbide-5-mononitrate (isosorbide mononitrate, ISMN, Mono Mack, CAS 16051-77-7) (40 mg ISMN in the morning, n = 187 vs. 20 mg ISMN b.i.d. morning and early afternoon, n = 195) were evaluated in an open, randomised study in patients with symptomatic myocardial ischemia. Circadian rate, frequency, severity and duration of angina pectoris attacks, as well as the additional need of short-acting nitrates were assessed on 3 consecutive days before and at the end of the first and second weeks of treatment, respectively. In both treatment groups, a statistically significant decrease of frequency and severity of angina pectoris attacks was observed as compared to baseline. Apart from minor variations, no statistically significant differences were found between the two treatment groups.


[Metabolic consequences of long-term antihypertensive treatment with co-dergocrine mesylate/nifedipine in high age groups]. Possible metabolic changes during chronic treatment is of specific importance for the development of antihypertensive drugs. The impact of a combination treatment with co-dergocrine mesilate/nifedipine (Pontuc; CAS 8067-24-1 resp. CAS 21829-25-4) on the lipid metabolism as well as hematological and biochemical values was evaluated in a group of hypertensive patients. HDL-cholesterol remained unchanged, whereas total cholesterol and LDL-cholesterol slightly decreased, apoprotein AI increased and apoprotein B decreased. In conclusion the results of this study indicate that co-dergocrine mesilate/nifedipine has no negative impact on the lipid metabolism in patients with essential hypertension.


Synthesis and inhibitory effects of 1,2,4-triazole derivatives on platelet aggregation. Various derivatives of triazole substituted in the 2-position were prepared, and their activity on platelet aggregation tested. Compounds 4 and 14 had the most powerful action. These agents were thought to inhibit platelet aggregation via an inhibition of the cyclo-oxygenase-peroxidase complex (PGS complex), preventing synthesis of prostaglandins.


Effect of anethole dithiolthione on human platelet aggregation. Anethole dithiolthione (ADT) (10 mumol/l) inhibited platelet aggregation and the formation of thromboxane (Tx)B2 in plasma in response to adenosine diphosphate (ADP), epinephrine and arachidonic acid (AA). ADT partially inhibited platelet aggregation and TxB2 formation in plasma induced by thrombin, phorbol myristate acetate and calcium ionophore A23187 and increased the lag time of collagen-induced aggregation at concentrations in the range 10-40 mumol/l. ADT (100 mumol/l) completely inhibited the aggregation of washed platelets challenged with thrombin. ADT had no additive effect on the inhibition of thrombin-induced platelet aggregation by acetylsalicylic acid. ADT was a more effective inhibitor of AA-induced platelet aggregation than butylated hydroxytoluene. ADT inhibited the release of 3H-AA from platelet phospholipids in response to ADP and collagen. It is suggested that ADT inhibits platelet aggregation by inhibiting thromboxane synthesis and preventing AA release.


Effects of the anti-platelet agent cilostazol on peripheral vascular disease in patients with diabetes mellitus. Effects of cilostazol (OPC-13013, CAS 73963-72-1), a selective inhibitor of platelet cAMP-phosphodiesterase, on peripheral vascular disease in diabetes mellitus were studied. Cilostazol in a dose of 200 to 300 mg/d was administered to 5 diabetic patients with arteriosclerosis obliterans. Skin temperature of the finger and the toe, which reflects blood flow to the tissue, was selected as an objective index of cilostazol effects and measured by infra-red thermography at a constant temperature of 26 degrees C. Before administration, digital skin temperatures were low in 9 limbs of 5 patients. 200 mg/d of cilostazol significantly (p less than 0.001) increased the digital skin temperatures of 8 limbs, the increase (mean +/- SD) ranging from 29.9 +/- 1.4 degrees C to 33.2 degrees C +/- 1.2 degrees C for the average skin temperatures and from 28.7 +/- 2.1 degrees C to 33.1 +/- 1.5 degrees C for the lowest ones. An increase in the dose to 300 mg/d resulted in further elevation of skin temperatures of the digits. Cilostazol constantly elicited an increase in blood flow to the digits within the range of its therapeutic dose. This effect was observed about 1 month after initiation of administration and persisted while administration was continued. The measurement of digital skin temperatures by infrared thermography provided a noninvasive means to individualize the dosage of cilostazol and to monitor the cilostazol effect and patient complicance during long-term administration. It is concluded that cilostazol exerts a potent and steady vasodilatory effect on peripheral circulation in patients with diabetes mellitus.


[Binding affinity of ibuprofen in human tissues]. In vitro binding of ibuprofen (CAS 15687-27-1) to various human tissues was studied to explain differences in tissue concentration after local application of ibuprofen cream. Radioactive ibuprofen, was incubated with human skin, subcutaneous tissue, muscle, tendon, joint capsule for 2 h at 37 degrees C. Tissue specimens were washed and radioactivity was measured by a liquid scintillation counter. The results show specific binding of ibuprofen to the various tissues of different degrees. Despite of interindividual differences muscle tissues showed the highest, tendons the lowest concentrations of ibuprofen. These findings may explain the in vivo results, where higher concentrations of ibuprofen were found in muscle tissue compared with subcutaneous tissue.


Study of the anti-inflammatory activity of Populus tremula, Solidago virgaurea and Fraxinus excelsior. Aqueous/alcoholic extracts of Populus tremula, Solidago virgaurea and Fraxinus excelsior (components of Phytodolor N) were tested individually and in 3 different combinations for anti-inflammatory activity using carrageenan induced edema and/or adjuvant induced arthritis of the rat paw. The tested combinations as well as the individual extracts significantly reduced the paw edema to varying degrees and also dose dependently inhibited the arthritic paw volume. The anti-inflammatory activity of the combinations was respectively comparable to the tested doses of diclofenac.


Effect of polyunsaturated phosphatidylcholine on liver regeneration onset after hepatectomy in the rat. The purpose of this study was to examine the effect of polyunsaturated phosphatidylcholine (PPC, Essentiale) on the onset of liver regeneration after partial hepatectomy (PH). Hepatectomy was carried out on rats, and immediately after operation doses of 25, 50 or 250 mg PPC/kg b.w. were injected into the vena femoralis. The control group consisted of saline-injected PH rats. The rats were killed 18, 21, 24 and 30 h after PH. The course of liver regeneration was assessed by measuring the incorporation of labelled thymidine into DNA and the hepatocyte mitotic activity. After PPC injection no values of liver DNA specific activity differing from baseline were observed. However, higher values of hepatocyte mitotic activity and lower liver triglyceride levels were found as compared to the saline-treated rats. These observations indicate that the administration of PPC has a favourable effect on liver regeneration after PH.


Therapeutic efficacy of latoconazole in formulations of clinical use on experimental dermatophytosis in guinea pigs. Therapeutic efficacy of clinical dosage forms of latoconazole (NND-318, CAS 101530-10-3), related compound of ketene dithioacetals with an imidazole ring, was examined on two experimental tinea models: a recently developed tinea pedis model and a conventional tinea corporis model in guinea pigs. The efficacy of the dosage forms was estimated on the basis of the rate of fungus-positive skin cultures and/or the score of skin lesions, and was compared to the other antifungal agents, bifonazole, clotrimazole and tolnaftate. While these reference agents exhibited curative effect on the tinea corporis model, the tinea pedis model was considerably resistant to the therapeutic treatment of the agents. The cream preparation and solution of lactoconazole at concentrations of more than 0.25% were highly effective in either tinea models, and at concentrations of more than 1%, lactoconazole achieved complete mycological cure. These results suggest that 1% of lactonazole is an optimal concentration for clinical use.


Fraction of absorbed drug at the time of peak plasma concentration in an open one-compartment model after simultaneous oral and intravenous administration. When a drug is simultaneously given orally and intravenously, the plasma concentration generally reaches a maximum. The purpose of this paper is to assess the extent of absorbed drug at the time Tmax.


Thrombolytic properties in a rabbit jugular vein thrombosis model of a tissue-type plasminogen activator mutant lacking the growth factor--and kringle one-domains. In a rabbit jugular vein thrombosis model, the thrombolytic properties of melanoma tissue-type plasminogen activator (mt-PA), recombinant t-PA (rt-PA), and a t-PA mutant that lacked the growth factor- and kringle one-domain (designated FK2P, as it contained the finger-, kringle two-, and protease-domain) were compared. Over the dose range tested (3.75 to 30 nmol/kg) a similar, dose-dependent, thrombolytic efficacy was found for the 3 enzymes when given as bolus injections. When given as a 90-min infusion, FK2P was superior to rt-PA at the tested dose of 15 nmol/kg. The effects on systemic plasma fibrinogen, plasminogen and alpha 2-antiplasmin consumption were much larger for rt-PA and FK2P than for mt-PA. The analogue FK2P was cleared more slowly than mt-PA or rt-PA, despite a similar initial alpha half-life: the mean residence time of FK2P was about 10 times as large as that of mt-PA or rt-PA. A comparison with data obtained in vitro suggested that in vivo thrombolytic performance cannot easily be predicted from in vitro data. The efficacy of a thrombolytic agent will also depend on its clearance rate, and may also, within a chosen model, depend on the way it is administered.


Response of newly hatched chicks to inanition. Newly hatched chicks were kept for up to 48 hours without food and water and compared with a control group of chicks given access to food and water within six hours of hatching. The deprived chicks progressively lost body water and developed increases in plasma total protein concentration consistent with a decrease in plasma volume. They demonstrated a stronger motivation to drink and drank more when offered water, suggesting that they had become dehydrated. There was evidence, however, that normally access to food was necessary to initiate drinking. The deprived chicks continued to rely on fat metabolism to supply their energy needs whereas the fed birds showed rapid reductions in circulating levels of beta-hydroxybutyrate and increases in plasma glucose and liver glycogen concentrations, indicating a change to carbohydrate-orientated energy metabolism. The deprived chicks lost weight at an average of 0.14 g/hour and weighed 16.5 g less than the fed chicks after 48 hours.


Inhibitors of human neutrophil cathepsin G: structural and biochemical studies. The interaction of a series of sulfonate and phosphate esters derived from N-hydroxysuccinimide with human leukocyte cathepsin G was investigated. The synthesized compounds were found to be time-dependent inhibitors of the enzyme. The composite interplay of steric and electronic effects leads to the formation of acyl enzymes of variable stability, ultimately resulting in partial or full recovery of enzymatic activity. Compounds acting via phosphorylation of the active site serine inactivated the enzyme rapidly and irreversibly.


Symptom responses of female Vietnam veterans to Operation Desert Storm. This study examined the status of symptoms of posttraumatic stress disorder (PTSD) in a cohort of women after the onset of Operation Desert Storm. Seventy-six non-treatment-seeking Vietnam veterans were obtained from lists of those who recently had participated in other research projects conducted at the National Center for Post-Traumatic Stress Disorder. Before the onset of Operation Desert Storm, subjects had completed a set of psychometrically valid instruments measuring general psychological symptoms and PTSD symptoms (e.g., SCL-90-R, Mississippi Scale for Combat-Related Posttraumatic Stress Disorder). On the basis of the latter scale, subjects were divided into groups with and without PTSD symptoms. At the height of the military conflict, subjects were recontacted and asked to complete the SCL-90-R and the Veterans Update Form, a measure assessing changes in PTSD symptoms. Multivariate analyses indicated that while most female Vietnam veterans experienced some intensification of stress-related symptoms during Operation Desert Storm, those who had previously reported high levels of PTSD were significantly more susceptible to greater distress. Results of this survey indicate that female Vietnam veterans with prior wartime exposure are an at-risk population for the intensification of stress symptoms after the recurrence of a military conflict.


Issues concerning the sedation of children for radiation therapy. Radiation therapy is a commonly used modality to treat certain pediatric malignancies. To optimally treat a child's tumor while sparing normal, developing tissues from both short- and long-term side effects, the child must be in a fixed and reproducible position on the treatment table. Many children, especially those under age three, very anxious children, and those who are developmentally disabled, may require daily sedation for a course of radiotherapy. Issues concerning the use of sedation, types of drugs used, and the nurse's role in the sedation process are presented in this article.


Genomic 3' terminal sequence comparison of three isolates of rabbit haemorrhagic disease virus. Comparison of sequence data is necessary in older to investigate virus origins, identify features common to virulent strains, and characterize genomic organization within virus families. A virulent caliciviral disease of rabbits recently emerged in China. We have sequenced 1100 bases from the 3' ends of two independent European isolates of this virus, and compared these with previously determined calicivirus sequences. Rabbit caliciviruses were closely related, despite the different countries in which isolation was made. This supports the rapid spread of a new virus across Europe. The capsid protein sequences of these rabbit viruses differ markedly from those determined for feline calicivirus, but a hypothetical 3' open reading frame is relatively well conserved between the caliciviruses of these two different hosts and argues for a functional role.


Geostatistical and morphological methods applied to three-dimensional microscopy. Three-dimensional (3-D) images of osteocyte lacunae were examined on a confocal microscope. Both geostatistical and morphological processing techniques were used to improve and to analyse them. By a geostastical approach, this study aims at improving 3-D confocal images before any further image processing. Optimized linear filters, which take account of the second-order statistics and the 3-D structure of the data, allow for the removal of imperfections such as noise and/or blur due to the axial convolution, and interpolate voxels on a face-centred cubic grid from an initial cubic grid. An application of this technique to 3-D biological images is demonstrated. In a second step, a 3-D binary image is digitized and cleaned with 3-D morphological filters. The standard 3-D measurements cannot be applied in this case, since all osteocytes cut the border of the field. For this reason a 3-D Boolean model has been adjusted, from which it is possible to derive all useful information on the repartition and the morphology of the osteocytes.


Surface-bound optical probes monitor protein translocation and surface potential changes during the bacteriorhodopsin photocycle. Light-induced H+ release and reuptake as well as surface potential changes inherent in the bacterio-rhodopsin reaction cycle were measured between 10 degrees C and 50 degrees C. Signals of optical pH indicators covalently bound to Lys-129 at the extracellular surface of bacteriorhodopsin were compared with absorbance changes of probes residing in the aqueous bulk phase. Only surface-bound indicators monitor the kinetics of H+ ejection from bacteriorhodopsin and allow the correlation of the photocycle with the pumping cycle. During the L550----M412 transition the H+ appears at the extracellular surface of bacteriorhodopsin. Surface potential changes detected by bound fluorescein or by the potentiometric probe 4-[2-(di-n-butylamino)-6-naphthyl]vinyl-1-(3-sulfopropyl)pyridinium betaine (di-4-ANEPPS) occur in milliseconds concomitantly with the formation and decay of the N intermediate. pH indicators residing in the aqueous bulk phase reflect the transfer of H+ from the membrane surface into the bulk but do not probe the early events of H+ pumping. The observed retardation of H+ at the membrane surface for several hundred microseconds is of relevance for energy conversion of biological membranes powered by electrochemical H+ gradients.


Use of computers among occupational health nurses: results of a survey. 1. A survey of occupational health nurses revealed that they are using computers at work for a variety of activities, and find computers to be useful tools in their jobs. 2. Occupational health nurses who use computers at work differ from nonusers in level of education, experience, and salary. 3. Nurses who do not use computers at work are interested in learning more about computers, and are willing to and interested in attending seminars to gain additional knowledge in this area.


National Asthma Education Program Expert Panel report: guidelines for the diagnosis and management of asthma. 1. Asthma, a chronic lung disease that affects 10 million Americans, is receiving increased attention as a public health problem due to increased occurrence and number of reported deaths. 2. This article presents an overview of the National Asthma Education Program Expert Panel guidelines, specifically addressing the definition and pathophysiology of asthma, effective management and control of asthma, and special considerations in asthma management. 3. Implications of the Expert Panel guidelines for occupational health nurses are addressed.


Understanding corporate cultures: use of systems theory and situational analysis. 1. A company's culture is its heroes, values, myths, and symbols. In strong cultures, the lives and productivity of employees are shaped by where they work. 2. Understanding corporate cultures, generally, and the one in which one works, specifically, can help the occupational health nurse develop and implement intervention strategies specific to that company's culture. 3. A strong culture has a system of rules dictating employee behavior. In a less strong culture, the rules and definitions are less rigid, facilitating more flexibility and less predictability.


Program descriptions: a positive outcome from change. 1. Major organizational restructuring was the impetus for standardization of occupational health practice at three hospital sites. 2. A generic framework was devised to describe the key occupational health programs. 3. The process of developing these program descriptions yielded obvious quality assurance monitors. 4. Corporate change often is perceived as threatening. The development of a useful tool through cooperative team effort has produced a positive outcome.


Environmental hazards in the workplace: legal and safety considerations. Both the employer and the employee must work together to provide the best possible work environment. The occupational health nurse can facilitate the interaction and communication between these groups to protect the employer and the employee. The company must follow legal guidelines related to hazardous materials, and the employees must understand and adhere to the established policies and procedures developed for their safety. The nurse can help assure that each is protected.


[Piriform sinus fistulas as rare hypopharyngeal abnormalities, diagnosis and therapy]. We report on the history, clinical findings, diagnostic examinations and the therapy of three patients with a left piriform sinus fistula, probably a fourth pharyngeal pouch. Our patients show recurrent inflammation and abscesses of the left lateral neck. X-ray (gastrografin swallow) showed the fistula originating from the floor of the left piriform sinus. CT-scan was helpful in the previously operated area, although the fistula could be shown by ultrasonic examination. Recurrent abscesses of the lateral neck may be caused by sinus piriform fistula. It is important to search for this rare malformation of the lateral neck.


[Necrosis of the tongue in temporal arteritis. Case report]. Because of the rich blood supply ischemic necrosis of the tongue is extremely rare. But if it happens though, the reason is most often due to temporal arteritis. It predominantly occurs in elderly women, like in our case. The correct diagnosis should be established as soon as possible because high-dose cortisone therapy both relieves the patient's complaints and prevents life-threatening complications.


[Ossification of the stapedius tendon as a rare cause of conductive hearing loss]. Ossification of the stapedial tendon is a very rare cause of conductive hearing loss. In addition to a review of the literature, two further cases of unilateral hearing impairment are reported. In both cases, division of the ossified tendon led to a complete disappearance of the middle-ear problem. Apart from middle ear malformations, infections of the middle ear may also give rise to ossification of the stapedial tendon.


[Holmium:YAG infrared laser and UV-Excimer. Laser effects on oral mucosa]. The in-vivo tissue ablation characteristics of a pulsed infrared laser (Holmium:YAG, lambda = 1980) and a pulsed excimer laser (XeCl, lambda = 308 nm) were studied. The holmium laser energy was coupled into nylon fibres. The excimer laser energy was coupled into quartz fibers. Laser incisions were made on white rats at different sites of the tongue. Power settings of 400 mJ per 2.5 microseconds pulse at 5 pulses per second for the holmium:YAG laser and 250 mJ per 1 microsecond pulse at 20 pulses per second for the excimer laser were used. Thermal gradients following laser application were measured. Excellent control of depth of tissue ablation was noted for both types of lasers. The tissue response was examined by light microscopy. Histological studies revealed tissue defects with clean contours. The coagulative necrosis zone for the 400 mJ holmium:YAG injuries measured from 370 microns to 640 microns, compared with 40 microns to 160 microns with the 250 mJ UV-excimer injuries. Wound healing was studied over a six-week period. Histologically, wound closure was completed after four weeks showing re-epithelialization without scar formation for both types of laser. The in-vivo tissue ablation characteristics and the operation reliability of the two lasers were compared.


[Immunologic behavior of human tracheal transplants]. In this study we tried to investigate the antigenicity of human tracheal allografts. We found transplantation antigens in the mucosa and the mixed glands using monoclonal antibodies in an indirect immunoperoxidase method. However these antigens were able to be destroyed with a chemical preservation procedure. The diminution of the antigenicity could be also achieved in a tracheal transplant human recipient. These results show that the preserved trachea may be immunologically a suitable material for reconstruction in surgery of the trachea.


[Interposition connectors with heparin coating for vascular anastomosis of microsurgical tissue flaps]. The intraluminal surface of interposition connectos which are introduced for the quick vascular connection of microsurgical free flaps was coated by heparin. This heparin coating proved to be stable at the polyurethane wall until the intraluminal surface of the interposition connectors with a length of 15 mm was coated by a layer of endothelial cells. 90% of the experimental venous connections didn't show a thrombus formation and were open for the blood flow 10 days postoperatively. The experimental free flaps which were connected by this way had survived 21 days postoperatively and showed a regular healing.


[Medical ENT endoscopic studies in the iron and steel industry]. 461 chromium- and nickel-exposed steelworkers were otolaryngologically examined by endoscopy. Besides chronic inflammatory diseases of the ear, nose and paranasal sinuses, the clinical diagnosis of leukoplakia and hyperplastic laryngitis was made for 30 workers. In no case a lesion suspicious of cancer was found. Office staff showing no such occupational risk factors served as the control group. It was found that in both groups smoking has to be assumed as the decisive factor for the development of chronic inflammation and precancerous changes in the oral cavity and the larynx.


[Perioperative single dose prevention with cephalosporins in the ENT area. A prospective randomized study]. Perioperative systemic antibiotic cover is usually recommended in major head and neck surgery with the aim of preventing postoperative wound infection. The surgeon must administer the drug most suitable for the particular operation and patient at an adequate dosage and over an adequate period. On the other hand, he has an obligation to minimize costs and find ways of avoiding the development of bacterial resistance to the antibiotic. This means that prophylactic antibiotics may not have too broad an antibacterial spectrum and may only be used over the short term. In an attempt to improve antibiotic prophylactic in head and neck surgery, we tested the effectiveness of a perioperative single-dose antibiotic cover with cefuroxim (Zinacef) in patients undergoing parotidectomy, sinus surgery or neck dissection with no transcutaneous exploration of the pharynx. This single-shot prophylaxis was compared with a three-shot, 24-hour regimen using the same antibiotic. The two regimens were equally effective in preventing wound infection and no case of infection was observed among the 106 patients involved. Preoperative concentrations of cefuroxim were measured in serum and tissue, and the large majority of specimens showed effective concentrations against the typical wound infection bacteria. In conclusion, a perioperative single-dose administration of cefuroxim proved to be suitable prophylaxis against postoperative wound infection in head and neck surgery.


[Dose-finding study for hyperfractionated, accelerated radiotherapy plus simultaneous carboplatin administration in patients with locoregionally advanced head and neck cancer]. Therapy of large inoperable squamous cell-carcinomas of the head and neck is considerably problemful. Combined simultaneous radio-chemotherapy with carboplatin and radiotherapy with hyperfractionated accelerated radiation is small effective than conventional monofractionated radiotherapy alone and may reduce side effects. Aim of the following study was dosage optimisation of the applicated carboplatin.


An objective structured clinical examination for the licentiate: report of the pilot project of the Medical Council of Canada. The Medical Council of Canada (MCC) administers a qualifying examination for the issuance of a license to practice medicine. To date, this examination does not test the clinical skills of history taking, physical examination, and communication. The MCC is implementing an objective structured clinical examination (OSCE) to test these skills in October 1992. A pilot examination was developed to test the feasibility, reliability, and validity of running a multisite, two-form, four-hour, 20-station OSCE for national licensure. In February 1991, 240 volunteer first- and second-year residents were tested at four sites. The candidates were randomly assigned to one of two forms of the test and one of two sites for two of the four sites. Generalizability analysis revealed that the variance due to form was 0.0 and that due to site was .16 compared with a total variance of 280.86. The reliabilities (inter-station) were .56 and .60 for the two forms. Station total-test score correlations, used to measure station validity, were significant for 38 of the 40 stations used (range .14-.60). The results of the OSCE correlated moderately with the MCC qualifying examination; these correlations were .32 and .35 for the two test forms. Content validity was assessed by postexamination questionnaires given to the physician examiners using a scale of 0 (low) to 10 (high). The physicians' mean ratings were: importance of the stations, 8.1 (SD, 1.8); success of the examination in testing core skills, 8.1 (SD, 1.6); and degree of challenge, 7.8 (SD, 2.1). The results indicate that a full-scale national administration of an OSCE for licensure is feasible using the model developed. Aspects of validity have been established and strategies to augment reliability have been developed.


A new program to enhance the teaching and assessment of clinical skills in the People's Republic of China. The authors describe the history and current state of development of an ongoing and still-evolving collaboration--between three Chinese medical schools, the China Medical Board of New York, Inc. (CMB), and one of the authors (PLS) and her staff at the University of Massachusetts Medical School--to enhance the teaching and assessment of clinical skills in the schools. The program grew out of a request by the three schools to the CMB for assistance in improving their process of evaluating students. Its goal is to develop a uniform and comprehensive course for teaching and assessing basic clinical skills. Standardized patients are also being used as instructors and evaluators. The program is co-sponsored by the three schools and the CMB. Background is given about the CMB, its involvement in Chinese medical education, and its American perspective on the nature of education in Chinese institutions of modern medicine and the factors that can assist and impede educational change. The authors present the mechanisms of the program, the benefits that have occurred so far, and the problems encountered in creating the model curriculum to teach clinical skills that is now being implemented by the three schools. Future plans are outlined, including the hope that the three schools will serve as a resource and disseminate some or all aspects of the new approach to similar medical schools in China, and that the program can serve as a model for implementation of similar programs in other countries.


Storytelling as a method for teaching values and attitudes. Storytelling is not widely accepted as a teaching method in medical education, sometimes for valid reasons that are explained by the authors. Yet clinician-teachers who choose and tell stories appropriately--especially if these are stories of their own clinical experiences--can stimulate their students to examine their values and attitudes in ways that would be hard or impossible to achieve by other methods. The present article, which contains a story of the type advocated, shows how storytelling can help students and residents discuss and overcome their crises of professionalization and come to grips with the troubling aspects of the doctor-patient relationship. The authors maintain that storytelling allows educators to bring the discussion of values and attitudes to where students are most likely to appreciate and understand the message--the clinical encounter.


Fourth-of-July fireworks over national health care. On July 3, President George Bush devoted his annual Fourth-of-July radio message to the American people to the issue of health care reform. What follows are slightly edited transcripts of the president's message and the response of the Democratic Party, which was delivered by Senate Majority Leader George Mitchell (D-Maine) and also broadcast on July 3.


What we say and what we do: self-reported teaching behavior versus performances in written simulations among medical school faculty. Much of the research in medical education is performed through the use of self-reporting questionnaires. Although this can be a valid way to conduct research, little effort has been made to examine the fit between what teachers self-report and what they actually do. In an attempt to investigate this, the authors interviewed 47 preclinical faculty members at two large medical schools in 1990. The faculty members were initially given a questionnaire asking them to self-report in four categories: interactive skills; knowledge or abilities they considered important for students to develop; factors that influenced their curriculum development; and sources from which they sought pedagogical assistance. This was followed by four written simulations that examined four areas of teaching: small-group discussions, course design, lecturing, and test construction. In addition, the authors specifically sought to identify any differences in teaching philosophy and practice between those preclinical faculty who were physicians and those who were not, as well as any interinstitutional differences. Although in certain instances there was a strong correlation between self-reporting and performance as measured by a simulated teaching scenario, in most instances the correlation was quite low. Consequently, the authors suggest that researchers must be careful in their use of self-reporting as a means of assessing teaching behavior, and that whenever possible, researchers should observe the teachers they are studying.


Conceptualizing curriculum for graduate medical education. Several recent developments affecting graduate medical education (GME) have kindled an interest in curriculum. For the most part, however, GME curriculum is being conceived in terms of behavioral learning objectives. The authors find this approach to curriculum ill-suited for the reality and complexity of housestaff training. Several other approaches are considered but none, they conclude, fits well with the mission of GME. Instead, they propose a more comprehensive experiential conception of curriculum for GME. This approach stems from an experiential learning paradigm and a commitment to curriculum as an expression of valued activities rather than of predetermined objectives. Taking as an example a curriculum for an ambulatory care block rotation, the authors show how an experiential curriculum can be developed and how it can be used to frame the residents' rotation, including patient care and didactic program.


Using a database to analyze core basic science content in a problem-based curriculum. As medical schools critically reevaluate their methods of instruction and as the number of innovative programs increases, the content delivered across disciplines must be carefully reviewed. However, few methods of content analysis have been applied to problem-based programs. In 1989-90 and 1990-91, the authors analyzed the distribution of basic science content in the 53 cases in the problem-based curriculum of Rush Medical College of Rush University. They developed a content vocabulary and created a database using a widely available computer software program. The content areas specific to each case were identified by faculty using the content vocabulary. To determine whether these content areas were actually identified by the students participating in the problem-solving sessions, the authors surveyed the 36 student participants in the classes of 1993 and 1994 and also interviewed the 15 faculty facilitators of the sessions. The surveys and interviews demonstrated that over 90% of the content areas identified by the faculty were actually covered by the students. The authors conclude that the database assists in their review of the curriculum for omission and redundancy. Other uses and limitations of this method are also discussed.


The role of natural science in the premedical curriculum. The authors examined the role of natural sciences in entering students' academic preparation for admission to the University of California, Los Angeles (UCLA), School of Medicine by analyzing the college transcripts of 290 students in the classes of 1990 and 1995 to determine the breadth of the students' college course work. In addition, during 1990 the authors surveyed 126 full-time basic science and clinical faculty about their views regarding the optimal requirements for admission. The vast majority of the students' college courses had been in the natural sciences, with no significant change in this finding over the five-year period. Of the faculty surveyed, 97 (77%) responded, stating that the majority of prerequisites should be in the natural sciences; their prerequisites were similar to UCLA's 1991-92 admission requirements. The authors suggest that the persistently narrow focus on the natural sciences as a preparation for medical school may explain the nationwide need to instruct medical students in the humanities, and that perhaps it is time to require non-science courses to broaden the educational backgrounds of entering students.


Women physicians in dual-physician relationships compared with those in other dual-career relationships. This study compared the career and domestic responsibilities of women physicians whose domestic partners were physicians (WP-Ps) with those of women physicians whose domestic partners were not physicians (WP-NPs). In 1988 the authors surveyed 602 women physicians in a large midwestern city regarding their career and domestic roles; 390 were physicians in training (students and residents), and 212 were physicians in practice (academic medicine and private practice). Overall, 382 (63%) responded; of the 382, 247 (65%) had domestic partners; of these 247, 91 (37%) were WP-Ps and 156 (63%) were WP-NPs. The WP-Ps were found to be twice as likely as the WP-NPs to interrupt their careers to accommodate their partners' careers. The WP-Ps also assumed significantly more domestic responsibilities and worked fewer hours practicing medicine than did the WP-NPs. The 163 women physicians in training (44-48%-of the WP-Ps and 119-76%-of the WP-NPs) demonstrated a more egalitarian division of labor overall, with no significant differences between the WP-Ps and the WP-NPs. The authors recommend that longitudinal studies be undertaken to determine whether women physicians in training continue this trend as they enter the practice of medicine.


Use of free time in the third and fourth years of a flexible curriculum. In 1988, the author surveyed senior students on their use of the six months of free time in the junior-senior biennial at the University of Minnesota Medical School-Minneapolis. The students were asked both to provide detailed reports on their free-time activities and to rank the three activities they considered to be most helpful to their professional development. Of 274 seniors, 191 (70%) responded. The students had used a majority of their free time for pursuits related to education and advancement of their careers. Approximately one-third of the time had been used for vacations. The activities the students viewed as most helpful included research, interviewing for residency positions, vacation, study elsewhere, and preparation for the National Board of Medical Examiners Part II examination. The author suggests that when free time is made available in a flexible curriculum, students use it to advantage in an educationally responsible manner.


Consultative and educational roles of a physical therapist in a family practice residency. Many medical schools and primary care residencies provide little formal instruction in musculoskeletal examination skills. Physical therapists are well qualified by knowledge and experience to teach these skills to students and residents. The authors present educational objectives that have been utilized since May 1988 in the physical therapy teaching portion of a sports medicine rotation in a family practice residency, and they describe the educational and consultative roles of a physical therapist in the residency. The therapist demonstrated the portion of the musculoskeletal examination that is important for the differentiation of contractile and inert tissue involvement, and she reviewed problems unique to athletes. The therapist's presence exposed students, residents, and faculty to the appropriate use of physical therapists as consultants and clinicians.


Alzheimer's disease: a model from the quantitative study of a large kindred. In an Italian kindred (family N), early onset Alzheimer's disease has been transmitted in a Mendelian autosomal fashion since the early 18th century. The age at death of affected members of the family varies widely, and was taken as an index of the age of expression, a measure of phenotypic variability. Either a gamma or a log-normal algorithm provides the best fit for the age at death distribution. Subsets of family N widely different as to time and place have the same age at death of patients: Environment appears to play a negligible role in the expression of disease. Pairwise correlation between an affected parent and child is zero: The disease is monogenic (no major expression gene). The same stochastic distribution of age of expression, but with late onset, and after correction for death from other causes, is compatible with the epidemiology of Alzheimer's disease in general. Mendelian genetics is a possible model for Alzheimer's disease etiology.


[Local immunological phenomena in the digestive system of the child and food allergy]. Over the last few years food allergies have become a clinical problem of prime importance in children. It is clear that food allergy is most frequent in infants. Food macromolecules are absorbed at every age, but most evidently during the first months of life. Many facts show that the barrier of the gastro-intestinal tract against harmful agents present in the intestinal lumen is not completely developed in infants and especially in newborns. Deficiencies in the secretory system for TgA contribute to greater permeability of the intestine for allergenic macromolecules in early childhood.


[Allergy to heparin. Particular problems in the pregnant woman]. Oral anti-coagulants are always contra-indicated for pregnant women. Only anti-coagulants of the types of heparin or its derivatives may be used. However, some patients have an allergy to Heparin. In this article the correct action to take both for diagnosis and prevention is studied. We present 2 observations of heparin-allergic pregnant women with, in the first case a tentative addiction to heparin which created a check and in the second case there was success in a second pregnancy with low-molecular weight heparin.


Population-based dementia screening program in Kuopio: the effect of education, age, and sex on brief neuropsychological tests. A neuropsychological screening battery including the Mini-Mental State Examination and four other brief cognitive tests (Russell's Adaptation of the Visual Reproduction Test, Trail Making Test, Verbal Fluency Tests on letters and category, and the Buschke Selective Reminding Test) was administered to a randomly selected population sample of 403 subjects aged 68 to 77 years to evaluate the effect of education, age, and sex on test scores. The difference in neuropsychological screening tests between various education groups (3 years or less, 4 to 6 years, 7 years or more) was statistically highly significant, even after the adjustment for the effect of age. The subscores and total scores were lowest in the minimal education group on every neuropsychological test. Education correlated more strongly than age with all neuropsychological test scores and subscores. The effect of sex on test results was seen only in some subscores of brief neuropsychological tests but not in a single item of the Mini-Mental State Examination. On the basis of our results, the effects of education, age, and sex have to be evaluated before using brief neuropsychological tests in population-based dementia screening.


[Spatial and temporal variations of the quantities of pollen from Cupressaceae-Taxaceae collected in Marseilles]. Marseille has two sites (Centre and North) where airborne pollens are collected and counted. The most numerous pollens are from Cupressaceae Taxaceae. We compared the quantities collected with both pollen traps during February, March and April 1988 and 1989. Most of pollinization takes place in March. In Marseille's Centre, there were three times more trapped pollen during 1989 than in 1988. In 1989, there was 40% more pollen on northern site as compared with the other one. As indicated in 1989 with both sites, it suggests a circadian periodicity of Cupressaceae Taxaceae pollen: their highest count is usually between 9 and 11 hours a.m.


Subclinical dementia: relevance of cognitive symptoms and signs. In 1971 and 1972, a representative sample of 70-year-old people (n = 392) from a population study was rated with regard to psychiatric symptoms and signs. Subjects showing evidence of dementia of any degree, including slight and questionable, were excluded (n = 18). The remaining 374 subjects were followed longitudinally for 9 years. Thirty-eight subjects developed severe dementia during this period. They were compared to those who did not with regard to 19 reported and 17 observed items concerning general psychopathology, seven reported items concerning sleep, and a global rating of mental health. Five items correlated with the development of severe dementia: number of remembered dreams per week, latency of speech, speed of speech, difficulties in finding words, and memory for recent events. A stepwise logistic regression procedure demonstrated that a low frequency of remembered dreams, a reduced memory for recent events, and difficulties in finding words made independent contributions to the prediction of dementia. There were no differences with regard to affective items. There were no differences between the subjects who developed senile dementia of the Alzheimer type and those with multi-infarct dementia. The results thus support the hypothesis that symptoms pertaining to cognitive functions are the first to appear in the development of dementia.


Patient education programmes in obstructive airway disease. The Ingelheim Model for promoting health through patient education. Chronic obstructive airway diseases (COAD) can be regarded as one of the major health problems needing environmental actions and screening programs for early detection and intensive patient education programs to cope with the needs of tertiary prevention. On the basis of our epidemiological study focused on COAD carried out in FRG (sample size August 1988: 63,000 participants) a patient education program has been developed and evaluated. In cooperation with general practitioners and pneumologists the program has been installed at practice and community level. The need for a patient education program has been assessed during the three years of the PNEUMOBIL-Project. It is not just a matter of cutting costs, but to a large extent a matter of the wellbeing of the patients and of reducing side effects to a minimum. The objective of the project can be split into three dimensions: (1) The cognitive aspect. Here significant lack of knowledge has to be overcome. At this point it has to be stated clearly that at the present time the medical community is not able to solve this problem on their own. (2) The psychomotoric aspect. Here the competent use of medication has to be trained. (3) The emotional aspect. The patient has to be motivated and integrated into the therapeutic process in a way that his compliance contributes significantly. The didactical concept consists of modules that can be used in varying sequences according to the needs of the target audience.(ABSTRACT TRUNCATED AT 250 WORDS)


Utility of the sickness impact profile in Parkinson's disease. The Sickness Impact Profile (SIP) is a questionnaire consisting of 136 items grouped into 12 categories and two dimensions (physical and psychosocial). To characterize its utility in Parkinson's disease (PD), we administered the SIP to 44 consecutive clinic patients with PD. Compared to 44 age- and sex-matched control subjects, PD patients had their greatest dysfunction in the categories of mobility, communication, and home management. The two items that PD patients most commonly endorsed were, "I am having trouble writing or typing" (75%) and, "My sexual activity is decreased" (61%). In general, these treated PD patients had greater dysfunction in the psychosocial than physical dimensions. Two simple PD-specific scales correlated well with the physical dimension score but less so with the psychosocial dimension, suggesting that the SIP assesses more functional domains than the PD-specific scales used. The SIP holds some promise as a broad measure of functional status in PD patients.


ECT-induced interictal delirium in patients with a history of a CVA. Earlier works have documented a high incidence of affective disorders in patients with a history of a cerebrovascular accident (CVA). In general, electroconvulsive therapy (ECT) has been reported to be effective in treating depressed patients with a history of CVA. Recent works have shown that preexisting structural brain changes may predispose patients to develop interictal ECT-induced delirium. However, the incidence of ECT-induced interictal delirium in patients with a history of CVA has not been directly studied. In this pilot study, the authors examined the incidence of ECT-induced interictal delirium in 14 depressed CVA patients compared with 14 elderly depressed controls (without a history of CVA). Interestingly, the overall incidence of delirium was identical in both groups (28.5%). However, consistent with previous works, some patients who had had a recent CVA involving the caudate nucleus appeared more likely to develop delirium. Thirteen of the 14 depressed CVA patients (92%) showed a moderate to significant improvement in their depressive symptoms from ECT. None of the patients developed significant cardiac complications during ECT.


p53 gene mutations in human endometrial carcinoma. Although carcinoma of the uterine endometrium is the most frequently diagnosed malignancy of the female reproductive tract, the molecular genetic features of this tumor have yet to be described in significant detail. Since mutations of the p53 tumor suppressor gene are the single most common genetic alteration found in human malignancies, we examined the hypothesis that p53 mutations occur in human endometrial carcinoma. Sequencing analysis of exons 5-8 revealed point mutations in 3 of 21 (14%) tumors; one mutation was an unusual single-base insertion at codons 176-177, resulting in a premature stop codon, whereas the other two were CGG----TGG transitions at codon 248. Two of these tumors showed reduction to homozygosity at the p53 allele, but one tumor apparently retained heterozygosity. These data indicate that p53 mutations occur in human endometrial carcinoma, although relatively infrequently, and that loss of the normal p53 allele does not necessarily occur with point mutation of the p53 gene in this tumor type.


Characterization of the Syrian hamster c-Ha-ras gene and intron-D-exon transcript. The coding sequences as well as 5'- and 3'-flanking sequences of the Syrian hamster c-Ha-ras gene were deduced from cDNA clones derived from embryo fibroblast cell lines. Sequences of introns B, C, and D were obtained from genomic DNA after amplification by the polymerase chain reaction. Sequence comparisons with rat, mouse, and human c-Ha-ras genes revealed a high degree of homology. One of 12 cDNA clones contained intron-D-exon (IDX) sequences due to alternative splicing that would encode a p19 Ha-ras gene product. Conservation between species suggests a functional role for the IDX, possibly as a negative control of p21 Ha-ras expression.